Abstract | BACKGROUNDS: Septic acute kidney injury (AKI) is a severe illness in clinics. Enriching researches investigated the regulatory network of AKI during the past decades, evidences showed that circular RNAs ( circRNAs) were involved in the molecular mechanism of human AKI. However, the special responses remain largely elusive. Thus, the study aims to investigate the function of circ_0114427 in the progression of AKI. METHODS: The levels of circ_0114427, miR-495-3p and Tumour Necrosis Factor Receptor-Associated Factor 6 ( TRAF6) were both assessed by quantitative real-time polymerase chain reaction (qRT-PCR). In addition, lipopolysaccharide (LPS) was applied to establish AKI cell model, and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was carried out to determine the viability of LPS-induced HK-2 cells. The expression of TRAF6, B-cell lymphoma-2 (Bcl-2), Bcl2-associated X (Bax), cleave- caspase 3, caspase 3, total IκBα (t-IκBα), phospho-IκBα (p-IκBα), total p65 (t-p65) and phospho-p65 (p-p65) were all detected via western blot. The levels of IL-1β and TNF-α were identified by western blot and ELISA. What's more, cell apoptosis was measured by flow cytometry. Lastly, dual- luciferase reporter, RNA Immunoprecipitation (RIP) and RNA pull-down assays were employed to verify the relationships between miR-495-3p and circ_0114427 or TRAF6 in vitro. RESULTS: The level of miR-495-3p was remarkably restrained while circ_0114427 and TRAF6 levels were specially reinforced in AKI patient serum samples and LPS-induced HK-2 cells. Moreover, IL-1β and TNF-α were highly expressed in LPS-induced AKI cells. Functionally, circ_0114427 was a sponge of miR-495-3p, and circ_0114427 silence-mediated effects in LPS-induced HK-2 cells were partly ameliorated by the addition of miR-495-3p inhibitor. Moreover, TRAF6 was a target gene of miR-495-3p, and the inhibiting effect of miR-495-3p on cell apoptosis and inflammatory response was mitigated by TRAF6 overexpression. Mechanistically, the circ_0114427/miR-495-3p/ TRAF6 axis modulated cell apoptosis and inflammatory response via NF-κB/p65 signalling pathway in AKI. CONCLUSION: Circ_0114427 regulated cell apoptosis and inflammatory response through miR-495-3p/ TRAF6 axis via NF-κB/p65 signalling pathway, providing a novel mechanism in clinical treatment of AKI patients.HighlightsCirc_0114427 is upregulated in serum specimens from septic AKI patients and LPS-induced HK-2 cells.LPS treatment suppresses cell viability and promotes apoptosis and inflammation in HK-2 cells.Circ_0114427 knockdown ameliorates the effects of LPS on cell viability, apoptosis and inflammation in HK-2 cells.Circ_0114427 regulates LPS-induced HK-2 cell injury by regulating miR-495-3p/ TRAF6/NF-κB/p65 axis.
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Authors | Lei Xu, Hongxia Cao, Peng Xu, Mingxi Nie, Chun Zhao |
Journal | Autoimmunity
(Autoimmunity)
Vol. 55
Issue 1
Pg. 52-64
(Feb 2022)
ISSN: 1607-842X [Electronic] England |
PMID | 34730059
(Publication Type: Journal Article)
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Chemical References |
- Lipopolysaccharides
- MicroRNAs
- NF-kappa B
- TNF Receptor-Associated Factor 6
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Topics |
- Acute Kidney Injury
(genetics, pathology)
- Apoptosis
(genetics)
- Humans
- Lipopolysaccharides
(adverse effects)
- MicroRNAs
(metabolism)
- NF-kappa B
(metabolism)
- TNF Receptor-Associated Factor 6
(genetics, metabolism, pharmacology)
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